Table 5.
Bioanalytical method validation results summary.
| Analyte Name | GS-5734 | GS-441524 | GS-704277 |
|---|---|---|---|
| Internal Standard (IS) | GS-829143 | GS-828840 | GS-829466 |
| Analytical Method Type | LC-MS/MS | ||
| Extraction Method | Protein Precipitation | ||
| Sample Volume | 50 μL | ||
| QC Concentrations | 4, 12, 200, 1600, and 3200 ng/mL | 2, 6, 100, 800, and 1600 ng/mL | 2, 6, 100, 800, and 1600 ng/mL |
| Standard Curve Concentrations | 4, 8, 40, 80, 400, 800, 3600, and 4000 ng/mL | 2, 4, 20, 40, 200, 400, 1800, and 2000 ng/mL | 2, 4, 20, 40, 200, 400, 1800, and 2000 ng/mL |
| Lower Limit Of Quantitation | 4 ng/mL | 2 ng/mL | 2 ng/mL |
| Upper Limit Of Quantitation | 4000 ng/mL | 2000 ng/mL | 2000 ng/mL |
| Mean Recovery of Analyte (%) | 97.4 | 105.4 | 81.1 |
| Mean Recovery of IS(%) | NA | ||
| LLOQ QC Intraday Precision Range (%CV) | 4.9 to 7.3 | 7.0 to 9.3 | 4.2 to 10.4 |
| LLOQ QC Intraday Accuracy Range (%RE) | −2.5 to 9.5 | −8.0 to 5.5 | −17.5 to −9.0 |
| Analytical QC Intraday Precision Range (%CV) | 1.2 to 4.8 | 2.3 to 6.6 | 1.6 to 4.6 |
| Analytical QC Intraday Accuracy Range (%RE) | −1.6 to 11.5 | −1.9 to 10.0 | −10.3 to −1.9 |
| LLOQ QC Interday Precision (%CV) | 7.7 | 9.8 | 9.2 |
| LLOQ QC Interday Accuracy (%RE) | 2 | 0 | −12.5 |
| Analytical QC Interday Precision Range (%CV) | 2.3 to 3.8 | 3.5 to 5.3 | 2.1 to 3.8 |
| Analytical QC Interday Accuracy Range (%RE) | 0.0 to 9.5 | −0.6 to 8.0 | −9.8 to −3.5 |
| Dilution Integrity | 10000 ng/mL diluted 20-fold | 5000 ng/mL diluted 20-fold | 5000 ng/mL diluted 20-fold |
| Selectivity | ≤20.0% LLOQ for analyte; ≤ 5.0% for IS | ≤20.0% LLOQ for analyte; ≤ 5.0% for IS | ≤20.0% LLOQ for analyte; ≤ 5.0% for IS |
| FA-Treated 2% Hemolyzed Plasma Test | No impact on assay performance | ||
| FA-Treated 5% Hemolyzed Plasma Test | No impact on assay performance | ||
| Lipemic Plasma Test | No impact on assay performance | ||